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[1]王乐,李晶,曹瑜,等.播娘蒿转录因子DsCBF与CRT/DRE元件的相互作用[J].应用与环境生物学报,2010,16(05):609-612.[doi:10.3724/SP.J.1145.2010.00609]
 WANG Le,LI Jing,CAO Yu,et al.Interaction of DsCBF of Descurainia sophia with CRT/DRE Element[J].Chinese Journal of Applied & Environmental Biology,2010,16(05):609-612.[doi:10.3724/SP.J.1145.2010.00609]
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播娘蒿转录因子DsCBF与CRT/DRE元件的相互作用()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
16卷
期数:
2010年05期
页码:
609-612
栏目:
胁迫抗性生理及分子机制专栏
出版日期:
2010-10-25

文章信息/Info

Title:
Interaction of DsCBF of Descurainia sophia with CRT/DRE Element
文章编号:
201004058
作者:
王乐李晶曹瑜张玲巨艳乔代蓉曹毅
四川大学生命科学学院,微生物与代谢工程四川省重点实验室 成都 610064
Author(s):
WANG LeLI JingCAO YuZHANG LinJU YanQIAO DairongCAO Yi
(Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu 610064, China)
关键词:
播娘蒿CBFCRT/DREEMSA银染
Keywords:
Descurainia sophia (L. ) Webb CBF CRT/DRE EMSA silver staining
分类号:
Q949.748.305 : Q78
DOI:
10.3724/SP.J.1145.2010.00609
文献标志码:
A
摘要:
CBF(C-repeat binding factor)是调控植物冷驯化相关基因表达的一种调控转录激活因子,CBF蛋白AP2结合domain可与COR基因启动子CBT/DRE元件特异性地结合,启动耐寒基因COR的表达. 为研究播娘蒿DsCBF的功能,构建pET32a-DsCBF原核表达载体,通过冻融法转化入大肠杆菌BL21(DE3)中,进行原核表达,并通过镍亲和层析纯化CBF蛋白. 利用EMSA(Electrophoretic mobility shift assay)分析DsCBF蛋白与播娘蒿DsCOR基因的启动子上CRT/DRE元件的相互作用. EMSA结果显示,DsCBF蛋白与含有CCGAC核心序列及TATA-box和AT-TA回文结构的50 bp探针结合有滞后现象,表明与DsCOR启动子上的CRT/DRE元件有特异性结合;而与仅有CCGAC的CRT/DRE核心序列的40 bp探针结合则无滞后现象,表明DsCBF与蛋白结合的识别可能与TATA-box和AT-TA回文结构有关. 图6 参19
Abstract:
CBF, the DNA-binding proteins of AP2/EREBP family, recognizes the cold- and dehydration- responsive DNA regulatory element, which is called CRT (C-repeat)/DRE (Dehydration-responsive element) for short. A prokaryotic expression vector pET32a-DsCBF was constructed to study the function of DsCBF in Descurainia sophia. The recombinant plasmid was transformed into E. coli BL21, and the fusion protein was further purified through Ni-affinity chromatography. CBF had an AP2 domain which was bound with the element of CRT/DRE specially. The transaction of CBF and promoter of COR were studied through EMSA (Electrophoretic mobility shift assay). Two probes were synthesized, and one of them had a TATA-box and AT-TA palindrome structure except core sequence CCGAC and the other only had the core sequence CCGAC. Our conclusion indicates that DsCBF bound with CRT/DRE element may be affected by TATA-box and AT-TA palindrome structure. Fig 6, Ref 19

参考文献/References:

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备注/Memo

备注/Memo:
国家自然科学基金项目(Nos. 30771312,30871321,30971817)资助 Supported by the National Natural Science Foundation of China (Nos. 30771312, 30871321 and 30971817)
更新日期/Last Update: 2010-10-25